Isolation and characterization of heparin from human mastocytoma tissue.
Polysaccharide was isolated from human spleen mastocytoma by proteolytic digestion, precipitation with cetylpyridinium chloride, digestion with chondroitinase ABC, and ion-exchange chromatography on DEAE-cellulose. The final product (0.7 mg per g of starting material, MW 8000) behaved like standard heparin on ion-exchange chromatography and on electrophoresis, and contained D-glucuronic acid, L-iduronic acid, D-glucosamine and sulfate in the proportions expected for heparin. Affinity chromatography on antithrombin-Sepharose separated a distinct high-affinity fraction (4-5% of the total material). Structural analysis of this fraction showed that about 10% of the D-glucosamine residues were N-acetylated, the remainder N-sulfated. The anticoagulant activity of the isolated heparin was 71 B.P. units per mg (whole-blood system), or 30 units per mg (anti-thrombin and chromogenic substrate). 205 and 10-15 units per mg (chromogenic assay) were found for high and low affinity fractions, respectively. These results demonstrate conclusively the occurrence of heparin in a human tissue.
D002846:Chromatography, Affinity; D002850:Chromatography, Gel; D002852:Chromatography, Ion Exchange; D005260:Female; D006493:Heparin / Q000032:analysis / Q000302:isolation & purification*; D006801:Humans; D008407:Mast Cells / Q000648:ultrastructure; D012515:Mast-Cell Sarcoma / Q000032:analysis* / Q000648:ultrastructure; D008854:Microscopy, Electron; D008875:Middle Aged; D013160:Splenic Neoplasms / Q000032:analysis* / Q000648:ultrastructure
D006493:Heparin